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  • 1
    Electronic Resource
    Electronic Resource
    Efficient enzymatic synthesis of 13 C,15N-labeled DNA for NMR studies (1997)
    Springer
    Journal of biomolecular NMR 10 (1997), S. 245-253 
    Details
    ISSN: 1573-5001
    Keywords: DNA ; DNA–protein complex ; Deoxythymidylate kinase ; Enzymatic synthesis ; Heteronuclear NMR ; Isotope labeling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The power of heteronuclear NMR spectroscopy to study macromoleculesand their complexes has been amply demonstrated over the last decade. Theobstacle to routinely applying these techniques to the study of DNA has beenthe synthesis of 13C,15N-labeled DNA. Here wepresent a simple and efficient method to generate isotope-labeled DNA forNMR studies that is as easy as that for isotope labeling of RNA. The methodwas used to synthesize a uniformly13 C,15N-labeled 32-nucleotide DNA that binds tohuman basic fibroblast growth factor with high affinity and specificity.Isotope-edited experiments were applied to the13 C,15N-labeled DNA bound to unlabeled protein,and the 13 C,15N-labeled DNA was also examined incomplex with 15N-labeled protein. The NMR experiments showthat the DNA adopts a well-defined stable structure when bound to theprotein, and illustrate the potential of13 C,15N-labeled DNA for structural studies ofDNA–protein complexes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1018358602001
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  • 2
    Electronic Resource
    Electronic Resource
    Cloning and expression of a Xenopus gene that prevents mitotic catastrophe in fission yeast (1995)
    Springer
    Molecular genetics and genomics 246 (1995), S. 387-396 
    Details
    ISSN: 1617-4623
    Keywords: Xenopus ovary ; cDNA expression library Yeast transformation ; Weel kinase ; Cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In fission yeast the Weel kinase and the functionally redundant Mikl kinase provide a regulatory mechanism to ensure that mitosis is initiated only after the completion of DNA synthesis. Yeast in which both Weel and Mik1 kinases are defective exhibit a mitotic catastrophe phenotype, presumably due to premature entry into mitosis. Because of the functional conservation of cell cycle control elements, the expression of a vertebrate weel or mikl homolog would be expected to rescue such lethal mutations in yeast. A Xenopus total ovary cDNA library was constructed in a fission yeast expression vector and used to transform a yeast temperature-dependent mitotic catastrophe mutant defective in both weel and mikl. Here we report the identification of a Xenopus cDNA clone that can rescue several different yeast mitotic catastrophe mutants defective in Weel kinase function. The expression of this clone in a weel/mikl-deficient mutant causes an elongated cell phenotype under non-permissive growth conditions. The 2.0 kb cDNA clone contains an open reading frame of 1263 nucleotides, encoding a predicted 47 kDa protein. Bacterially expressed recombinant protein was used to raise a polyclonal antibody, which specifically recognizes a 47 kDa protein from Xenopus oocyte nuclei, suggesting the gene encodes a nuclear protein in Xenopus. The ability of this cDNA to complement mitotic catastrophe mutations is independent of Weel kinase activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00288613
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    A frequency-dependent finite-difference time-domain formulation for induced current calculations in human beings (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Bioelectromagnetics 13 (1992), S. 543-555 
    Details
    ISSN: 0197-8462
    Keywords: broadband irradiation ; short pulses ; tissue dispersion ; induced currents ; SARs ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: The finite-difference time-domain (FDTD) method has been used to calculate SARs and induced currents involving whole-body or partial-body exposures of models to spatially uniform or nonuniform (far-field or near-field), to sinusoidally varying EM fields, or to transient fields such as those associated with electromagnetic pulses. However, a weakness of the FDTD algorithm is that the dispersion of the tissue's dielectric properties is ignored and frequency-independent properties are assumed. Although this is permissible for continuous-wave or narrow-band irradiation, the results may be highly erroneous for short pulses, in which ultra-wide bandwidths are involved. In some recent publications, procedures are described for one- and two-dimensional problems for media in which the complex permittivity ∊ * (ω) may be described by a single-order Debye relaxation equation or a modified version thereof. These procedures based on a convolution integral describing D(t) in terms of E(t) cannot be extended to human tissues for which multiterm Debye relaxation equations must generally be used.We describe here a new differential-equation approach that can be used for general dispersive media. We illustrate the use of this approach by one- and three-dimensional examples of media for which ∊ * (ω) is given by a multiterm Debye equation, and for an approximate two-thirds muscle-equivalent model of the human body. Based on a single run involving a Gaussian pulse, the frequency-dependent FDTD [(FD)2TD] method allows calculations of SARs and induced currents at various frequencies by taking the Fourier components of the induced E fields. The (FD)2TD method can also be used to calculate coupling of the short (ultra-wideband) pulses to the human body. 1992 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bem.2250130609
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    Paper (German National Licenses)
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